Quantitative measurement of ultraviolet-induced damage in cellular DNA by an enzyme immunodot assay

Biotechnol Appl Biochem. 1989 Feb;11(1):11-7. doi: 10.1111/j.1470-8744.1989.tb00047.x.

Abstract

A simple enzyme immunoassay procedure was developed for the quantitative determination of 254-nm uv-induced DNA damage in cells. With the use of specific antibodies to uv-irradiated DNA and horseradish peroxidase-conjugated antibody to rabbit IgG, the extent of damaged DNA in uv-irradiated rat spleen mononuclear cells was quantitatively measurable. Through the use of this method, the amount of damaged DNA present in 2 X 10(5) cells irradiated at a dose of 75 J/m2 was estimated to be 7 ng equivalents of the standard uv-irradiated DNA. In addition, when the cells, irradiated at 750 J/m2, were incubated for 1 h, the antigenic activity of DNA decreased by 40%, suggesting that a repair of the damaged sites in DNA had proceeded to some extent in the cells.

MeSH terms

  • Animals
  • DNA Damage*
  • Densitometry
  • Immunoenzyme Techniques
  • Rats
  • Rats, Inbred Strains
  • Spleen / cytology
  • Spleen / radiation effects
  • Ultraviolet Rays / adverse effects*